Cryptococcus-Specific Methods
General Molecular Biology Methods
Cell storage
Bacterial transformation - chemical
Cell wall transfer assay
DNA blotting
DH & DC assays
E. coli competent cells
Genomic DNA -
large
&
small scale
preps
PCR colony screening
Growth media:
basic
,
specialty
,
sporulation
, and
capsule-inducing
Restriction enzyme digestion tips
Immunofluorescence -
anti-capsule antibody
&
other antibodies
RNA blotting
Isolation of GXM
Site directed mutagenesis (by kit)
RNA preparation
TA cloning
Transformation - biolistics,
electroporation
Lab Methods
General Biochemistry Methods
Antibody purification
Beta-glucuronidase assay
Antibody tagging (Cy3)
Biorad protein assay (
regular
&
DC
)
Capsule transfer assay
Immunoblotting
Cell wall tagging (fluorescein)
Pouring protein gels (
regular
&
gradient
)
Membrane preparations -
small
,
medium
, &
large scale
Purification of His-tagged proteins from bacteria (small scale)
Thin layer chromatography
Staining protein gels (
Coommassie
&
silver
)
TCA precipitation
Equipment
AKTA chromatography system
Lyophilizer
Mini-ultracentrifuge
Scintillation counter
